利用表达谱芯片数据筛选不同表达丰度的内参基因

Screening of reference genes of different abundance using gene expression microarrays

目的基于表达谱芯片的检测结果,筛选多个内参基因,用于小家鼠肝脏组织中具有不同表达丰度基因的定量检测。方法应用表达谱芯片技术,完成小鼠肝脏组织的表达谱检测;依据基因表达丰度将基因分为3组,并进一步通过变异系数(coefficient of variation,CV)组内筛选候选内参基因;采用实时荧光定量PCR技术(realtime quantitative polymerase chain reaction,q PCR)和ge Norm软件确定内参基因。结果表达谱芯片成功采集超过60000个小鼠肝脏组织中转录本的表达量数据,并将之分为低、中、高3个组合。最终筛选了低表达Casp2和Lrrc14、中表达Nrd1和Trpc4ap、高表达Atp5a1和Clu,共6个内参基因。结论基于表达谱芯片数据筛选的6个内参基因,可适用于q PCR技术准确定量小家鼠肝组织转录组中不同表达丰度基因的表达量。

Objective Based on the data detected using gene expression microarray,to select multiple reference genes and use them to quantify transcriptome genes of different abundance in the mouse liver tissue. Methods To detect global transcriptome genes in the mouse liver tissues using gene expression microarray. All the genes were sorted into different groups according to their expression level,followed by coefficient of variation（ CV） analysis. Real-time quantitative polymerase chain reaction（ q PCR） and ge Norm software were used to further identify the reference genes. Results The expression levels of over 60,000 genes were obtained from microarray screening,and divided them into low,moderate and high expression groups. Finally six reference genes were screened,i. e. Casp2 and Lrrc14 as low abundance,Nrd1 and Trpc4 ap as moderate abundance,and Atp5a1 and Clu as high abundance genes. Conclusions The six reference genes derived from microarray data can be used to accurately quantify the global transcriptome genes with various expression levels.